Agent for the treatment of infections

ABSTRACT

PCT No. PCT/JP95/00491 Sec. 371 Date Sep. 18, 1996 Sec. 102(e) Date Sep. 18, 1996 PCT Filed Mar. 17, 1995 PCT Pub. No. WO95/25517 PCT Pub. Date Sep. 28, 1995The present invention relates an agent for the treatment of infections, which comprises as the active ingredient at least one member selected from the group consisting of aconite-alkaloids, aconite tuber and an extract thereof, gingerol and analogues thereof, and rhizomes of ginger and a substance therefrom. The agent of the present invention has significant infection-protective and recovery effects and is useful to treat and prevent a variety of infections such as virus infections, fungal infections, opportunistic infections, etc.

This application is a 371 of PCT/JP95/00491 filed Mar. 17, 1995.

TECHNICAL FIELD

The present invention relates to an agent for the treatment ofinfections, which is effective against virus infections, fungalinfections, etc.

BACKGROUND ART

It is said that the infections have drastically changed in recent years.The classical infections have become latent, while infections that aredifficult to treat and occur depending on the resistance of a patient toinfections are increasing. Such infections occurring depending on thestate of a patient are called opportunistic infections. Typicalpathogens for opportunistic infections (herpes simplex virus type I) andtheir diseases are shown in Table 1. Many of the diseases are seriousalthough they are caused by the same kind of pathogen.

                  TABLE 1    ______________________________________                            infection under decreased    first infection                relapse     infection-protective ability    ______________________________________    stomatitis  lip herpes  herpes eczema                corneal herpes                            generalized infection                            adult encephalitis    ______________________________________

The opportunistic infections refer to the state in which infections havebeen induced by pathogens of usually little pathogenity because theinfection-protective ability of a patient is decreased for some reasons(increased susceptibility to infections). Hence, an agent having arecovery effect of infection-protective ability and an antimicrobialagent effective against pathogens are used to treat the opportunisticinfections. In particular, the agent having a recovery effect ofinfection-protective ability is regarded as indispensable for essentialtreatment of the opportunistic infections.

However, the opportunistic infections show similar clinical symptomsalthough many are caused by a plurality of pathogens, and from theclinical symptoms, it is difficult to identify the pathogen, so it isalso difficult to select a suitable antimicrobial agent for use in theopportunistic infections. Even if a suitable antimicrobial agent can beselected, substantial treatment of the opportunistic infections wouldnot be achieved until the resistance of the patient is recovered.However, there are not a few cases where the lowered general conditionsof the patient permit such an agent having a recovery effect ofinfection-protective ability to adversely induce a decrease in theinfection-protective ability of the patient. At present, there is noagent that can be administered with confidence to treat or prevent theopportunistic infections.

Aconite tuber is tuberous roots of Aconitum carmichaeli of the familyRanunculaceae or other plants of the same genus, and it has been usedfor a long time as cardiotonic, analgesic, antiphlogistic, etc.

Aconite-alkaloids are aconite tuber-derived alkaloids known to haveefficacy as analgesic, antiphlogistic, etc., and typical examples arebenzoylmesaconine, benzoylaconine, benzoylhypaconine, 14-anisoylaconine,neoline, ignavine, mesaconine, hypaconine, 16-epi-pyromesaconitine,16-epi-pyraconitine, 15-α-hydroxyneoline and ajaconine.

Rhizomes of Zingiber officinale Roscoe of the family Zingiberaceae have,for a long time, been used as ginger in medical formulation regarded aspeptic, anti-emetic or analgesic, or in other medical formulation. Inaddition, gingerol as a component of ginger is known to have efficacy asantipyretic, analgesic, etc.

However, aconite tuber or an extract thereof or aconite-alkaloids orderivatives thereof, or ginger, dried ginger or an extract thereof, orgingerol or analogues thereof have never been used to treat infectionssuch as virus infections, fungal infections, opportunistic infections,etc.

DISCLOSURE OF INVENTION

The object of the present invention is to provide an agent for thetreatment of infections which has significant recovery effect ofinfection-protective ability.

The present inventors searched for a pharmaceutical preparation toimprove the infection-protective ability of a patient that is decreasedfor some reasons (increased susceptibility to infections), and thepresent inventors found that aconite-alkaloids, a plant containing thesame or an extract thereof and aconite-alkaloid derivatives, as well asgingerol, a plant containing the same or a substance therefrom andgingerol analogue have inhibitory action on increased susceptibility toinfections, and thereby they completed the present invention.

That is, the present invention encompasses:

(1) An agent for the treatment of infections, which comprises as theactive ingredient a compound represented by the general formula (I):##STR1## wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³,R¹⁴, and R¹⁵ are the same or different and independently represent ahydrogen atom, hydroxyl group, substituted or unsubstituted C₁ -C₇ alkylgroup, substituted or unsubstituted C₂ -C₇ alkenyl group, substituted orunsubstituted C₂ -C₇ alkynyl group, substituted or unsubstituted C₃ -C₇cycloalkyl group, substituted or unsubstituted C₄ -C₇ cycloalkenylgroup, substituted or unsubstituted acyl group, substituted orunsubstituted acyloxy group, substituted or unsubstituted acyloxy-C₁ -C₇alkyl group, substituted or unsubstituted C₂ -C₇ alkoxycarbonyl group,substituted or unsubstituted C₂ -C₇ alkenyl-oxycarbonyl group,substituted or unsubstituted aryloxycarbonyl group, substituted orunsubstituted C₁ -C₇ alkoxy group, substituted or unsubstituted C₂ -C₇alkenyloxy group, substituted or unsubstituted C₂ -C₇ alkynyloxy group,substituted or unsubstituted C₃ -C₇ cycloalkyloxy group, substituted orunsubstituted C₄ -C₇ cycloalkenyloxy group, substituted or unsubstitutedaryloxy group, substituted or unsubstituted aryl-C₁ -C₇ alkyloxy group,substituted or unsubstituted aryl-C₂ -C₇ alkenyloxy group, substitutedor unsubstituted C₁ -C₇ alkoxy-C₁ -C₇ alkyl group or substituted orunsubstituted C₁ -C₇ alkoxy-C₁ -C₇ alkoxy group, or R² and R³ maytogether represent an epoxy group, R¹ and R¹⁴ may together represent anepoxy group, and R⁷ and R⁸ may together represent an oxo group, or apharmaceutically acceptable salt thereof.

(2) An agent for the treatment of infections, which comprises as theactive ingredient a compound represented by the general formula (II):##STR2## wherein R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹, R²², R²³, A¹, A², A³, A⁴and A⁵ are the same or different and independently represent a hydrogenatom, hydroxyl group, substituted or unsubstituted C₁ -C₇ alkyl group,substituted or unsubstituted C₂ -C₇ alkenyl group, substituted orunsubstituted C₂ -C₇ alkynyl group, substituted or unsubstituted C₃ -C₇cycloalkyl group, substituted or unsubstituted C₄ -C₇ cycloalkenylgroup, substituted or unsubstituted acyl group, substituted orunsubstituted acyloxy group, substituted or unsubstituted acyloxy-C₁ -C₇alkyl group, substituted or unsubstituted C₂ -C₇ alkoxycarbonyl group,substituted or unsubstituted C₂ -C₇ alkenyl-oxycarbonyl group,substituted or unsubstituted aryloxycarbonyl group, substituted orunsubstituted C₁ -C₇ alkoxy group, substituted or unsubstituted C₂ -C₇alkenyloxy group, substituted or unsubstituted C₂ -C₇ alkynyloxy group,substituted or unsubstituted C₃ -C₇ cycloalkyloxy group, substituted orunsubstituted C₄ -C₇ cycloalkenyloxy group, substituted or unsubstitutedaryloxy group, substituted or unsubstituted aryl-C₁ -C₇ alkyloxy group,substituted or unsubstituted aryl-C₂ -C₇ alkenyloxy group, substitutedor unsubstituted C₁ -C₇ alkoxy-C₁ -C₇ alkyl group or substituted orunsubstituted C₁ -C₇ alkoxy-C₁ -C₇ alkoxy group, A⁶ represents ahydrogen atom, and R²⁰ and R²¹ and/or R²³ and A³ may together representan oxo group, A¹ and A⁵ and/or A³ and A⁴ may together represent a singlebond, A² and A⁵ may together represent an epoxy group, and A⁴ and A⁶ maytogether represent an ethyleneoxy group, or a pharmaceuticallyacceptable salt thereof.

(3) An agent for the treatment of infections, which comprises aconitetuber or an extract thereof as the active ingredient.

(4) An agent for the treatment of infections, which comprises as theactive ingredient a compound represented by the general formula (III):

    Ar-CH.sub.2 CH.sub.2 CO-R.sup.24                           (III)

wherein Ar represents a substituted or unsubstituted phenyl group, R²⁴represents a substituted or unsubstituted C₁ -C₇ alkyl group,substituted or unsubstituted C₂ -C₇ alkenyl group, substituted orunsubstituted aryl-C₁ -C₇ alkyl group or substituted or unsubstitutedaryl-C₂ -C₇ alkenyl group.

(5) An agent for the treatment of infections, which comprises rhizomesof ginger or a substance therefrom as the active ingredient.

In the present specification, the C₁ -C₇ alkyl group includes forexample a methyl group, ethyl group, n-propyl group, isopropyl group,n-butyl group, isobutyl group, sec-butyl group, t-butyl group, pentylgroup, hexyl group, and heptyl group. The C₂ -C₇ alkenyl group includesfor example a vinyl group, allyl group, propene-2-yl group,but-1-en-4-yl group, and but-2-en-2-yl group. The C₂ -C₇ alkynyl groupincludes for example an ethynyl group and propargyl group. The C₃ -C₇cycloalkyl group includes for example a cyclopentyl group and cyclohexylgroup. The C₄ -C₇ cycloalkenyl group includes for example acyclopent-2-enyl group and cyclohex-3-enyl group. The acyl groupincludes for example an aliphatic acyl group composed of theaforementioned C₁ -C₇ alkyl group, C₂ -C₇ alkenyl group, C₂ -C₇ alkynylgroup, C₃ -C₇ cycloalkyl group or C₄ -C₇ cycloalkenyl group; an aromaticacyl group composed of an aryl group such as phenyl group, naphthylgroup, pyrrolyl group, pyrazolyl group, imidazolyl group, triazolylgroup, tetrazolyl group, furyl group, thienyl group, and pyridyl group,examples being a benzoyl group and anisoyl group; and aryl-C₁ -C₇alkyl-CO-- group and aryl-C₂ -C₇ alkenyl-CO-- group composed of theaforementioned aryl group and of the aforementioned C₁ -C₇ alkyl groupor C₂ -C₇ alkenyl group. The acyloxy group includes an acyloxy groupcomposed of the aforementioned acyl group, and examples are C₁ -C₇alkyl-COO--, C₂ -C₇ alkenyl-COO--, C₂ -C₇ alkynyl-COO--, C₃ -C₇cycloalkyl-COO--, C₄ -C₇ cycloalkenyl- COO--, aryl-COO--, aryl-C₁ -C₇alkyl-COO--, and aryl-C₂ -C₇ alkenyl-COO--. The acyloxy-C₁ -C₇, alkylgroup includes an acyloxyalkyl group composed of the aforementionedacyloxy group and C₁ -C₇ alkyl group, an example is a benzoyloxymethylgroup. The C₂ -C₇ alkoxycarbonyl group includes an alkoxycarbonyl groupcomposed of the aforementioned C₁ -C₇ alkyl group. The C₂ -C₇alkenyl-oxycarbonyl group includes an alkenyloxycarbonyl group composedof the aforementioned C₂ -C₇ alkenyl group. The aryloxycarbonyl groupincludes an aryloxycarbonyl group composed of the aforementioned arylgroup. The C₁ -C₇ alkoxy group includes an alkoxy group composed of theaforementioned C₁ -C₇ alkyl group. The C₂ -C₇ alkenyloxy group includesan alkenylalkoxy group composed of the aforementioned C₂ -C₇ alkenylgroup. The C₂ -C₇ alkynyloxy group includes an alkynylalkoxy groupcomposed of the aforementioned C2-C₇ alkynyl group. The C₃ -C₇cycloalkyloxy group includes a cycloalkyloxy group composed of theaforementioned C₃ -C₇ cycloalkyl group. The C₄ -C₇ cycloalkenyloxy groupincludes a cycloalkenyloxy group composed of the aforementioned C₄ -C₇cycloalkenyl group. The aryloxy group includes an aryloxy group composedof the aforementioned aryl group. The aryl-C₁ -C₇ alkyloxy groupincludes an arylalkyloxy group composed of the aforementioned aryl groupand C₁ -C₇ alkyl group. The aryl-C₂ -C₇ alkenyloxy group includes anarylalkenyloxy group composed of the aforementioned aryl group and C₂-C₇ alkenyl group. The C₁ -C₇ alkoxy-C₁ -C₇ alkyl group includes analkoxyalkyl group composed of the aforementioned C₁ -C₇ alkoxy group andC₁ -C₇ alkyl group, and an example is a methoxymethyl group. The C₁ -C₇alkoxy-C₁ -C₇ alkoxy group includes an alkoxyalkoxy group composed ofthe aforementioned C₁ -C₇ alkoxy group, and an example is amethoxymethoxy group.

The aforementioned substituent groups may be substituted with a hydroxylgroup, halogen atom, nitro group, substituted or unsubstituted aminogroup, substituted or unsubstituted succinimido group, etc.

As R²⁴ in the above formula (III), the aryl-C₁ -C₇ alkyl group includesan aralkyl group composed of the aforementioned aryl group and C₁ -C₇alkyl group, examples being benzyl group and phenethyl group, and thearyl-C₂ -C₇ alkenyl group includes an arylalkenyl group composed of theaforementioned aryl group and C₂ -C7 alkenyl group, an example being astyryl group.

The active ingredients of the agent of the present invention are notparticularly limited insofar as they contain aconite-alkaloids. Examplesof active ingredients are aconite-alkaloids such as benzoylmesaconine,benzoylaconine, benzoylhypaconine, 14-anisoyl- aconine, neoline,mesaconine, hypaconine, 16-epi-pyromesaconitine, 16-epi-pyraconitine,15- α-hydroxyneoline, moticamine, moticoline, lappaconine, Excelsine,delvestidine, N-acetyldelectine, ajacine, anhweidelphinine,methyllycaconitine, avadharidine, septentrionine, andersonine,ajaconine, dihydroajaconine, ignavine, septentriosine, spiradine-C,spiramine-C and spirasine-III, or aconite tuber which is a plantcontaining them or an extract thereof. A crude drug containing anaconite-alkaloid other than aconite tuber itself or an extract thereofmay also be used. Further, a mixed crude drug containing aconite tuberas a constituent crude drug or an extract thereof may also be used.

The aforementioned aconite-alkaloids are known compounds and can beobtained in conventional processes.

The structures of typical aconite-alkaloids are shown below: ##STR3##

It is further possible to make use of esterified, etherified or acylatedderivatives prepared for example in the following manner: ##STR4##wherein Bz stands for benzoyl group, As for anisoyl group, MOM formethoxymethyl group, DIEA for N,N-di-isopropylethylamine, and Ac foracetyl group.

An extract from the above crude drug includes those extracted with avariety of aqueous solvents, preferably water. For example, the extractcan be obtained by extracting the crude drug with a 10- to 20- foldvolume of hot water and filtering the extract. If necessary, the extractmay be dried for use as dried powder.

As the active ingredients of the present agent, rhizomes of ginger and asubstance therefrom may be used in any form such as ginger, dried gingeror an extract thereof. A mixed crude drug containing ginger or driedginger as a constituent crude drug or an extract thereof may also beused.

The above extract includes those extracted with a variety of aqueoussolvents, preferably water. For example, the extract can be obtained byextracting the crude drug with a 10- to 20-fold volume of hot water andfiltering the extract. If necessary, the extract can be dried for use asdried powder.

The preparation of an aconite tuber extract as one of the activeingredients of the present agent is illustrated by reference to thefollowing specific examples.

SPECIFIC EXAMPLE 1

400 ml of distilled water was added to 20 g of aconite tuber and boiledto evaporate half the volume. The extract was filtered and lyophilizedto give 5.5 g of dried extract.

SPECIFIC EXAMPLE 2

After addition of 4 L of purified water, 200 g of aconite tuber wasextracted by heating for 1 hour at 100° C. The extract was centrifugedto separate the residue. The supernatant was filtered through a 0.3 μ mmembrane filter (Toyo Roshi Co., Ltd.) to remove bacteria. Thetransparent filtrate was ultrafiltrated at a pressure of 3 kg/cm²through Diafilter G-10T™ of 152 mm in diameter (Bioengineering Co.,Ltd.; a fractionation molecular weight of 10,000) attached to the bottomof a 2.0 L vessel while purified water was added as the solution becameconcentrated in the vessel. The ultrafiltrate was thus obtained.

As the active ingredients of the present agent, aconite-alkaloids suchas benzoylmesaconine, benzoylaconine, benzoylhypaconine,14-anisoylaconine, neoline, mesaconine, hypaconine,16-epi-pyromesaconitine, 16-epi-pyraconitine, ¹⁵ -a -hydroxyneoline,moticamine, moticoline, lappaconine, Excelsine, ajaconine,dihydroajaconine, ignavine, septentriosine, spiradine-C, spiramine-C andspirasine-III can be obtained by suitable extraction of aconite tuber(see for example Yakugaku Zasshi, 104 (8), 858-866 (1984)).

The preparation of an extract from dried ginger as an active ingredientof the present agent is illustrated by reference to the followingspecific examples.

SPECIFIC EXAMPLE 3

400 ml of distilled water was added to 20 g of dried ginger and boiledto evaporate half the volume. The extract was filtered and lyophilizedto give 3.5 g of dried extract.

SPECIFIC EXAMPLE 4

After addition of 4 L of purified water, 200 g of dried ginger wasextracted by heating for 1 hour at 100 ° C. The extract was centrifugedto separate the residue. The supernatant was filtered through a 0.3 μ mmembrane filter (Toyo Roshi Co., Ltd.) to remove bacteria. Thetransparent filtrate was ultrafiltrated at a pressure of 3 kg/cm²through Diafilter G-10T™ of 152 mm in diameter (Bioengineering Co.,Ltd.; a fractionation molecular weight of 10,000) attached to the bottomof a 2.0 L vessel while purified water was added as the solution becameconcentrated in the vessel. The ultrafiltrate was thus obtained.

As the active ingredient of the present agent, gingerol can be obtainedby suitable extraction of ginger or dried ginger (see for exampleAustralian Journal of Chemistry, 22, 1033 (1969)).

The infection-therapeutic action of the active ingredients of thepresent agent is described by reference to the following experimentalexamples.

EXPERIMENTAL EXAMPLE 1

The action of the active ingredients of the present agent was examinedin terms of suppressor cell activities induced in a model of increasedsusceptibility to infections (thermally injured mice) previouslyreported in FASEB Journal, 6, 1981 (1992) by the present inventors.

(1) Model of increased susceptibility to infection (thermally injuredmice)

8-week-old BALB/c male mice were anesthetized by intraperioneallyadministrating 0.8 mg/20 g of pentobarbital. The hair between the thighsand armpits of the mice was removed with a pair of hair clippers, andthird degree burns (about 30% of body area on 20 g mouse) were generatedon the mice by attaching the bared skin to an asbestos wire gauze (2×3cm) previously heated by a gas burner. Immediately after burns weregenerated, the mice were intraperioneally given 3 ml of physiologicalsaline as the model of increased susceptibility to infections (thermallyinjured mice). As normal mice, other mice were treated in the samemanner except that the operation of generating burns was not carriedout.

(2) Determination of survival rate

One day after burns were generated, the mice were infectedintraperioneally with 3×10³ PFU/kg of herpes simplex virus type I. Oneday before the infection, 1 day and 4 days after the infection, thethermally injured mice were orally given each of the active ingredientsof the present agent (4 mg/kg of aconite tuber extract or 1 μ g/kg ofaconite-alkaloid) through a stomach probe. Twenty-five days after theinfection, their survival rate was determined by comparison with thoseof the control groups, i.e. the thermally injured mice infected withherpes simplex virus type I and the normal mice infected with the samevirus.

                  TABLE 2    ______________________________________    active ingredient                   average survival days                                 survival rate (%)    ______________________________________    control group of infected                   11.8          0    thermally injured mice    control group of infected                   >24.4         90    normal mice    aconite tuber extract obtained                   >22.8         85    in Specific Example 1    benzoylmesaconine                   >21.5         85    benzoylaconine >22.5         80    benzoylhypaconine                   >22.4         80    14-anisoylaconine                   >22.5         80    neoline        >22.3         80    ignavine       >21.9         75    mesaconine     >23.1         85    hypaconine     >23.1         85    16-epi-pyromesaconitine                   >22.4         80    16-epi-pyraconitine                   >21.2         70    15-α-hydroxyneoline                   >20.9         70    ajaconine      >22.3         80    ______________________________________

(3) Measurement of contrasuppressor cell activities

Contrasuppressor cell activities, i.e. cells activities suppressing thesuppressor cell activities induced in thermally injured mice weredetermined in mixed lymphocyte tumor cell reaction (MLTR) by mixing andincubating the following cells for 5 days in a 96-wells round-bottommicroplate. Reaction cells: Spleen mononuclear cells (referred tohereinafter as SMNCs) (5.0×10⁴ cells/well) obtained from normal BALB/cmice. Stimulator cells: EL-4 tumor cells (5.0×10⁴ cells/well).Suppressor cells: SMNCs (2.5×10⁵ cells/well) obtained from the model ofincreased susceptibility to infections. Contrasuppressor cells: SMNCs(2.5×10⁵ cells/well) obtained from a model (BALB/c mice) of increasedsusceptibility to infections, previously given an active ingredient ofthe present agent (4 mg/kg of aconite tuber extract, or 1 μ g/kg ofaconite-alkaloid) twice every third day.

Before incubation, the cells except for the reaction cells were treatedwith 40 μ g/ml of mitomycin C solution at 37° C. for 30 minutes.Twenty-four hours before the conclusion of 3-days incubation, 0.5 μCi/well of ³ H-thymidine was added to the well, and the amount of ³ H-thymidine incorporated into the reaction cells was determined in aliquid scintillation counter to evaluate the contrasuppressor cellactivities induced against the suppressor cell activities byadministration of the active ingredient of the present agent.

The results are shown in Tables 3 and 4.

                  TABLE 3    ______________________________________                       incorporation of .sup.3 H-thymidine    active ingredient  (CPM ± SE)    ______________________________________    incubation of reaction cells and                       12413 ± 1438    stimulator cells (normal model)    incubation of reaction cells, stimulator                       2650 ± 70    cells and suppressor cells (model of    increased susceptibility to infection)    aconite tuber extract obtained in                       15888 ± 1775    Specific Example 1    ______________________________________

                  TABLE 4    ______________________________________                      incorporation of .sup.3 H-thymidine    active ingredient (CPM ± SE)    ______________________________________    incubation of reaction cells and                      12413 ± 1438    stimulator cell (normal model)    incubation of reaction cells stimulator                      2650 ± 70    cells and suppressor cells (model of    increased susceptibility to infection)    benzoylmesaconine 14754 ± 1832    benzoylaconine    16140 ± 1738    benzoylhypaconine 21319 ± 903    14-anisoylaconine 12720 ± 1578    neoline           13744 ± 526    ignavine          12831 ± 2255    mesaconine        15791 ± 2778    hypaconine        14111 ± 2232    16-epi-pyromesaconitine                      16891 ± 2232    16-epi-pyraconitine                      15654 ± 1209    15-α-hydroxyneoline                      14498 ± 1960    ajaconine         17655 ± 1879    ______________________________________

EXPERIMENTAL EXAMPLE 2

In this example, 7-week-old BALB/c mice were infected intraperioneallywith a lethal dose (3×10⁵ PFU/kg) of herpes simplex virus type I. Twodays before the infection, 1 day and 4 days after the infection, theactive ingredient of the present agent (10μ g/kg) or physiologicalsaline (control) was orally administered through a stomach probe intothe mice. Twenty-five days after the infection, their survival rate wasdetermined to evaluate the action of the active ingredient. In thisexperiment, the experimental mice were given the active ingredient ofthe present agent before and after infection with the virus to evaluateits action mainly on the prevention and treatment of the virusinfection.

The result of the extract from dried ginger obtained in Specific Example3 is shown in Table 5. The result of gingerol is shown in Table 6.

                  TABLE 5    ______________________________________    administered agent                average survival days                              survival rate (%)    ______________________________________    physiological                12.4          0    saline (control)    extract of dry ginger                >23.1         70    obtained in Specific    Example 3    ______________________________________

                  TABLE 6    ______________________________________    administered agent                average survival days                              survival rate (%)    ______________________________________    physiological                8.5           0    saline (control)    gingerol    >24.4         90    ______________________________________

EXPERIMENTAL EXAMPLE 3

Mice as opportunistic infection model inoculated intraperioneally with0.1 ml/mouse of a culture supernatant of AIDS virus (LP-BM5 virus)infected SC-1 cells were orally given benzoylmesaconine or physiologicalsaline. Then, the mice were observed for 150 days to determine theirsurvival rate. The results are shown in Table 7.

                  TABLE 7    ______________________________________    administered agent                   number of mice                              survival rate (%)    ______________________________________    physiological saline                   10         0    (0.2 ml/mouse)    benzoylmesaconine                   10         80    (1 μg/kg)    ______________________________________

The contrasuppressor cell activities induced against suppressor cellactivities by administration of benzoylmesaconine was evaluated in thesame manner as in "(3) Measurement of contrasuppressor cell activities"in Experimental Example 1. As suppressor cells, spleen mononuclear cellswere prepared from mice 110 days after infection with LP-BM5 virus. Theresults are shown in Table 8.

                  TABLE 8    ______________________________________               incorporation of .sup.3 H-thymidine                                suppressing rate*    administered agent               (CPM ± SE)    (%)    ______________________________________    physiological saline               1,730 ± 289   81    (0.2 ml/mouse)    benzoylmesaconine               8,826 ± 535   4    (1 μg/kg)    ______________________________________     *Suppressing rate =  (incorporation in normal model minus incorporation i     contrasuppressor cells)/(incorporation in normal model minus incorporatio     in model of increased susceptibility to infection)! × 100

EXPERIMENTAL EXAMPLE 4

Mice as opportunistic infection model 80 days after intraperionealinfection with 0.1 ml/mouse of a culture supernatant of AIDS virus(LP-BM5 virus) infected SC-1 cells were inoculated intraperioneally with1×10³ PFU/kg of herpes simplex virus type I (HSV). Then, the mice wereorally given benzoylmesaconine or physiological saline daily for 30 daysbeginning 60 days after the infection with the AIDS virus and observeddaily to determine their survival rate. The results are shown in FIG. 1.

EXPERIMENTAL EXAMPLE 5

Mice as opportunistic infection model 80 days after intraperionealinfection with 0.1 ml/mouse of a culture supernatant of AIDS virus(LP-BM5 virus) infected SC-1 cells were inoculated intravenously with3×10⁵ cells/mouse of Candida albicans. Then, the mice were orally givenbenzoylmesaconine or physiological saline daily for 30 days beginning 60days after the infection with the AIDS virus and observed daily todetermine their survival rate. The results are shown in FIG. 2.

EXPERIMENTAL EXAMPLE 6

Mice as opportunistic infection model inoculated intraperioneally with0.1 ml/mouse of a culture supernatant of AIDS virus (LP-BM5 virus)infected SC-1 cells were inoculated intravenously with 3×10⁵ cells/mouseof Candida albicans, and the effect of benzoylmesaconine by oraladministration was examined. The mice were slaughtered 2, 7 and 14 daysafter inoculation with Candida albicans, and the number of alivebacteria in the kidney was determined by the colony counting method. Theresults are shown in FIG. 3.

EXPERIMENTAL EXAMPLE 7

Thermally injured mice were infected intravenously with 2.5×10³PFU/mouse of cytomegalovirus (CMV), then given orally benzoylmesaconine(10 μ g/kg/day) or physiological saline (0.2 ml/mouse/day), and observedfor 25 days after the infection. The opportunistic infection of thethermally injured mice to CMV was efficiently protected byadministration of benzoylmesaconine. The results are shown in Table 9.

                  TABLE 9    ______________________________________                          average    rate (%)    administered agent               number of mice                          survival days                                     survival rate (%)    ______________________________________    physiological               10         12.4       0    saline (control)    benzoylmesaconine               10         >22.1      100    ______________________________________

EXPERIMENTAL EXAMPLE 8

SCID mice inoculated with 5×10⁶ cells/mouse of peripheral lymphocytes(suppressor cells) from patients with burns were infected with 2.5×10³PFU/mouse of CMV. Then, the mice were orally given benzoylmesaconine (10μ g/kg/day) or physiological saline (0.2 ml/mouse/day). As shown inTable 10, the results indicated the protective effect ofbenzoylmesaconine on CMV.

                  TABLE 10    ______________________________________    administered agent                   number of mice                              survival rate (%)    ______________________________________    physiological  10         0    saline (control)    benzoylmesaconine                   10         100    ______________________________________

EXPERIMENTAL EXAMPLE 9

Mice 1 day after burns generated as opportunistic infection model wereinfected with 2×10⁴ cells/mouse of Candida albicans and then givenintraperioneally benzoylmesaconine (1 μ g/kg) or physiological saline(0.2 ml/mouse). As a result, the resistance of the thermally injuredmice to the fungal infection was completely recovered (see FIG. 4).

EXPERIMENTAL EXAMPLE 10

Spleen mononuclear cells, total T cells, CD4⁺ T cells and CD8⁺ T cellswere prepared from spleens of mice as opportunistic infection model 80days after infection with LP-BM5 virus. The reaction cells, stimulatorcells and suspected suppressor cells were used in the ratio of 1:1:5,and the mixed lymphocyte tumor cell reaction (MLTR) was carried out inthe same manner as in "(3) Measurement of contrasuppressor cellactivities" in Experimental Example 1. As MLTR control, 5×10⁴ cells/wellof the reaction cells were incubated with only the stimulator cells. Theresults are shown in Table

                  TABLE 11    ______________________________________                 incorporation of .sup.3 H-thymidine                                  suppressing    cells used in MLTR                 (CPM ± SE)    rate* (%)    ______________________________________    MLTR control 15,330 ± 726  --    spleen mononuclear cells                 3,066 ± 122   80    total T cells                 2,759 ± 105   82    CD4.sup.+  T cells                 14,870 ± 873   3    CD8.sup.+  T cells                 2,300 ± 118   85    ______________________________________     *Suppressing rate =  (incorporation in MLTR control minus incorporation i     suppressor cells)/(incorporation in MLTR control)! × 100

In Table 11, the cells responsible for suppressor activities aresuggested to be CD8⁺ T cells.

When normal CD8³⁰ T cells and CD8⁺ suppressor T cells were incubated for24 to 72 hours under CO₂ in the absence of stimulation, interleukin 4and interleukin 10 activities were found in the culture.

Furthermore, thermally injured mice became resistant to Candida albicansinfection by replacement of their CD8⁺ suppressor T cells by normal CD8⁺T cells. On the other hand, the susceptibility of normal mice toinfections was increased by introduction of CD8⁺ suppressor T cells fromthermally injured mice, and their resistance to infections was recoveredby removal of the introduced CD8⁺ suppressor T cells with antibodies andsubsequent reconstitution with normal T cells. These results indicatethat CD8⁺ suppressor T cells act an important role in fungal infectionsas well.

From the foregoing results, it is evident that the active ingredients ofthe present agent have a significant effect on the inhibition ofincreased susceptibility to infections as well as therapeutic effects oninfections such as virus infections, fungal infections, etc. Hence, thepresent agent is useful to treat and prevent infections such as virusinfections, fungal infections, opportunistic infections, etc.

An active ingredient of the present agent, i.e. aconite tuber or anextract thereof can be used with confidence because it has a longhistory in itself or as a constituent crude drug of a Chinese herbalremedy and its safety was confirmed. This is also evidenced by the factthat every mouse and rat survived after oral administration of 10 g ofaconite tuber. The acute toxicities LD₅₀ (mg/kg)! of aconite- alkaloidssuch as benzoylmesaconine, benzoylaconine, benzoylhypaconine,14-anisoylaconine, neoline, ignavine, mesaconine, hypaconine,16-epi-pyromesaconitine, 16-epi-pyraconitine, ¹⁵ -α-hydroxyneoline andajaconine differ from one another but generally range from about 20 to500 mg/kg.

Another active ingredient of the present agent, i.e. ginger, driedginger or an extract thereof can also be used with confidence because ithas a long history in itself or as a constituent crude drug of a Chineseherbal remedy and its safety was confirmed as well. For example, theacute toxicity LD₅₀ (mg/kg)! of an extract of dried ginger (extractedwith water) was reported to be 33500 in terms of crude drug toxicity(Shoyakugaku Zasshi, 37 (1), 37-83 (1983)). The acute toxicity ofgingerol LD₅₀ (mg/kg)! is generally about 250 mg/kg (J. Pharm. Dyu.7,836-848 (1984)).

Hereinafter, the agent of the present invention will be furtherdescribed by reference to pharmaceutical manufacturing and dosage.

The present agent can be manufactured into pharmaceuticals by combiningthe aforementioned active ingredient with suitable pharmaceuticalcarriers. The agent can be administered in any form. For example, it canbe administered orally in the form of tablet, capsule, granule, finegranule, powder, etc., or parenterally in the form of suppository,injection, external preparation, etc.

In order to achieve the desired effect, the dose of the activeingredient depends on the weight and age of a patient and the degree ofdisease. The dose of the active ingredient per day, administered atintervals into an adult, is preferably about 0.5 to 2 g for aconitetuber, about 50 μ g to 5 mg for aconite-alkaloids, about 0.5 to 9 g forginger or dried ginger, and about 50μ g to 100 mg for gingerol.

A pharmaceutical preparation to be orally administrated is manufacturedin a usual manner using starch, lactose, sucrose, mannitol,carboxymethylcellulose, corn starch, inorganic salts, etc.

If necessary, this type of preparation can make use of binders,disintegrators, surfactants, lubricants, enhancers for the fluidity,flavoring agents, colorants, perfumes, etc. Examples are as follows:Binders!

Starch, dextrin, powdered acacia, gelatin, hydroxypropyl starch,methylcellulose, carboxymethylcellulose sodium, hydroxypropyl-cellulose,microcrystalline cellulose, ethylcellulose, polyvinylpyrrolidone,Macrogol. Disintegrators!

Starch, hydroxypropyl starch, carboxymethylcellulose sodium,carboxymethylcellulose calcium, carboxymethylcellulose, low-substitutedhydroxypropylcellulose. Surfactants!

Sodium lauryl sulfate, soybean lecithin, sucrose esters of fatty acid,polysorbate 80. Lubricants!

Talc, waxes, hydrogenated vegetable oils, sucrose esters of fatty acid,magnesium stearate, calcium stearate, aluminum stearate,poly(ethyleneglycol). Enhancers for the fluidity!

Light anhydrous silicic acid, dried aluminum hydroxide gel, syntheticaluminum silicate, magnesium silicate.

The present agent can also be administered in the form of suspension,emulsion, syrup or elixir, and it may contain corrigents and colorants.

Pharmaceutical preparations parenterally administrated, for exampleexternal preparations such as ointment, lotion, liniment, etc., aremanufactured in a usual manner generally using carriers (base materials)such as liquid paraffin, Iso-Par, vaseline, silicone oil, aliphatichigher alcohols (palmityl alcohol, oleyl alcohol), higher aliphaticacids (myristic acid, stearic acid), esters of fatty acid(microcrystalline wax, isopropyl myristate, etc.), lanolin, plastibase(a mixture of liquid paraffin and polyethylene), poly(ethyleneglycol),water, etc. Where required, it is possible to add emulsifiers (fattyacid monoglyceride, sorbitan ester of fatty acid, polyoxyethylene laurylether, etc.), wetting agents (glycerin, propylene glycol, sorbitol,etc.), antiseptics (methyl or propyl paraoxybenzoate etc.), antioxidants(BHA etc.), pH adjusting agent (citric acid etc.), suspending agents(CMC etc.) and other pharmaceuticals (itching-preventive agent,analgesic, etc.). Preparations to be percutaneously absorbed fall intothe aforementioned preparations.

For preparation of injections, use can be made of diluent that isgenerally distilled water for injection, physiological saline, anaqueous glucose solution, vegetable oils for injection, sesame oil,peanut oil, soybean oil, corn oil, poly(propyleneglycol),poly(ethyleneglycol), etc. If necessary, disinfectants, antiseptics andstabilizers may also be added. For the sake of stability, an injectionpreparation may be manufactured as a lyophilized sample etc. in vials sothat a liquid preparation is reconstituted just before use. If required,isotonic agents, stabilizers, disinfectants and soothing agents mayfurther be added.

In order to achieve the desired effect, the content of the activeingredient of the present agent in the preparation depends on the dosageform, the age of the patient and the degree of disease. The dose forexternal preparation is generally 0.01 to 5, preferably 0.1 to 0.5 μ gper gram of base material.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows the effect of benzoylmesaconine on the survival rate ofHSV-infected mice. In FIG. 1, the symbol◯ represents a group given 1 μg/kg/day of benzoylmesaconine, the symbol Δ represents a group given 0.1μ g/kg/day of benzoylmesaconine, and the symbol  represents a groupgiven physiological saline.

FIG. 2 shows the effect of benzoylmesaconine on the survival rate ofCandida albicans-infected mice. In FIG. 2, the symbol □ represents agroup given 1 μ g/kg/day of benzoylmesaconine, the symbol ▪ represents agroup given 10μ g/kg/day of benzoylmesaconine, and the symbol represents a group given physiological saline.

FIG. 3 shows the effect of benzoylmesaconine on the propagation ofCandida albicans in the kidney. In FIG. 3, the symbol represents a groupgiven 1 μ g/kg/day of benzoylmesaconine, and the symbol ▪ represents agroup not given benzoylmesaconine.

FIG. 4 shows the effect of benzoylmesaconine on the survival rate ofthermally injured mice infected with Candida albicans. In FIG. 4, thesymbol  represents a group given benzoylmesaconine, and the symbol ◯represents a group given physiological saline.

BEST MODE FOR CARRYING OUT THE INVENTION

The pharmaceutical manufacturing of the agent of the present inventionis described in more detail by reference to the following examples,which however are not intended to limit the present invention.

EXAMPLE 1

    ______________________________________    (1) Corn starch             21 g    (2) Microcrystalline cellulose                                10 g    (3) Carboxymethylcellulose calcium                                7 g    (4) Light anhydrous silicic acid                                1 g    (5) Magnesium stearate      1 g    (6) Dried extract of aconite tuber obtained                               160 g    in Specific Example 1    total                      200 g    ______________________________________

The ingredients (1) to (6) were uniformly mixed and compression-mouldedin a compression machine to give tablets (200 mg/tablet).

One tablet contains 160 mg of the dried extract of aconite tuberobtained in Specific Example 1, and 20-80 tablets are administered dailyinto an adult at suitable intervals.

EXAMPLE 2

    ______________________________________    (1) Corn starch           188 g    (2) Magnesium stearate     2 g    (3) Carboxymethylcellulose calcium                               8 g    (4) Light anhydrous silicic acid                               1 g    (5) Benzoylmesaconine      1 g    total                     200 g    ______________________________________

The ingredients (1) to (5) were uniformly mixed, compression-moulded ina compression machine, ground in a crusher and screened to givegranules.

One gram of the granules contains 5 mg of benzoylmesaconine and 0.5-5 gof the granules is administered daily into an adult at suitableintervals.

EXAMPLE 3

    ______________________________________    (1) Corn starch          198.5 g    (2) Light anhydrous silicic acid                             1 g    (3) 14-anisoylaconine    0.5 g    total                    200 g    ______________________________________

The ingredients (1) to (3) were uniformly mixed and 200 mg wasintroduced into No. 2 capsule.

One capsule contains 0.5 mg of 14-anisoylaconine and 1-4 capsules areadministered daily into an adult at suitable intervals.

EXAMPLE 4

300 g of alanine (pyrogen-free) was added to 20 L of a solutioncontaining the aconite tuber extract obtained in Specific Example 2, andit was dissolved and lyophilized. The lyophilized sample was introducedinto 900 vials to give an injection. This injection was of permeabilitysufficient to pass the pyrogen test in accordance with the JapanesePharmacopoeia.

EXAMPLE 5

    ______________________________________    (1) 16-epi-pyromesaconitine                            0.05 g    (2) Isopropyl myristate 5 g    (3) Plastibase          94.95 g    ______________________________________

The ingredients (1) and (2) were mixed and added gradually to theingredient (3) with stirring. The mixture was homogenized to give anoily ointment.

EXAMPLE 6

    ______________________________________    (1) 15-α-hydroxyneoline                            0.05 g    (2) Isopropyl myristate 5.95 g    (3) Isopropyl myristate 10 g    (4) Vaseline            66 g    (5) Liquid paraffin     5 g    (6) Microcrystalline wax                            13 g    ______________________________________

The ingredients (3) to (6) were molten by heating, and a mixture of (1)and (2) was added thereto at 45 to 50° C. It was homogenized withstirring until it was solidified to form an oily ointment.

EXAMPLE 7

    ______________________________________    (1) Ajaconine            0.05 g    (2) Poly(ethyleneglycol) (400)                             11.95 g    (3) Poly(ethyleneglycol) (400)                             12 g    (4) Poly(ethyleneglycol) (4000)                             76 g    ______________________________________

The ingredients (3) and (4) were molten at 70° C., and a mixture of (1)and (2) was added thereto at 50° C. It was homogenized with stirringuntil it was solidified to form a hydrophilic ointment. Carbol 934 ™ canfurther be mixed with the hydrophilic ointment.

EXAMPLE 8

    ______________________________________    (1) Corn starch            21 g    (2) Microcrystalline cellulose                               10 g    (3) Carboxymethylcellulose calcium                               7 g    (4) Light anhydrous silicic acid                               1 g    (5) Magnesium stearate     1 g    (6) Dried extract of dried ginger                              160 g    obtained in Specific Example 3    total                     200 g    ______________________________________

The ingredients (1) to (6) were uniformly mixed and compression-mouldedin a compression machine to give tablets (200 mg/tablet).

One tablet contains 160 mg of the dried extract of dried ginger obtainedin Specific Example 3, and 20-80 tablets are administered daily into anadult at suitable intervals.

EXAMPLE 9

    ______________________________________    (1) Corn starch           188 g    (2) Magnesium stearate     2 g    (3) Carboxymethylcellulose calcium                               8 g    (4) Light anhydrous silicic acid                               1 g    (5) Gingerol               1 g    total                     200 g    ______________________________________

The ingredients (1) to (5) were uniformly mixed, compression-moulded ina compression machine, ground in a crusher and screened to givegranules.

One gram of the granules contains 5 mg of gingerol and 1-10 g of thegranules is administered daily into an adult at suitable intervals.

EXAMPLE 10

    ______________________________________    (1) Corn starch           19 g    (2) Light anhydrous silicic acid                              1 g    (3) Dried extract of dried ginger    obtained in Specific Example 3                             180 g    total                    200 g    ______________________________________

The ingredients (1) to (3) were uniformly mixed, and 200 mg wasintroduced into No. 2 capsule.

One capsule contains 180 mg of the dried extract of dried ginger, and20-80 capsules are administered daily into an adult at suitableintervals.

EXAMPLE 11

300 g of alanine (pyrogen-free) was added to 20 L of a solutioncontaining the extract of dried ginger obtained in Specific Example 4,and it was dissolved and lyophilized. The lyophilized sample wasintroduced into 900 vials to give an injection. This injection was ofpermeability sufficient to pass the pyrogen test in accordance with theJapanese Pharmacopoeia.

EXAMPLE 12

    ______________________________________    (1) Dried extract of dried ginger                             0.05 g    obtained in Specific Example 3    (2) Isopropyl myristate  5 g    (3) Plastibase           94.95 g    ______________________________________

The ingredients (1) and (2) were mixed and added gradually to theingredient (3) with stirring. The mixture was homogenized to give anoily ointment.

EXAMPLE 13

    ______________________________________    (1) Gingerol            0.05 g    (2) Isopropyl myristate 5.95 g    (3) Isopropyl myristate 10 g    (4) Vaseline            66 g    (5) Liquid paraffin     5 g    (6) Microcrystalline wax                            13 g    ______________________________________

The ingredients (3) to (6) were molten by heating, and a mixture (1) and(2) were added thereto at 45 to 50° C. It was homogenized under stirringuntil it was solidified to form an oily ointment.

EXAMPLE 14

    ______________________________________    (1) Gingerol             0.05 g    (2) Poly(ethyleneglycol) (400)                             11.95 g    (3) Poly(ethyleneglycol) (400)                             12 g    (4) Poly(ethyleneglycol) (4000)                             76 g    ______________________________________

The ingredients (3) and (4) were molten at 70° C., and a mixture of (1)and (2) was added thereto at 50° C. It was homogenized with stirringuntil it was solidified to form a hydrophilic ointment. Carbol 934.sup.™can further be mixed with with the hydrophilic ointment.

INDUSTRIAL APPLICABILITY

The agent of the present invention has significant recovery effect ofinfection-protective ability, and it is useful to treat and preventvarious infections such as virus infections, fungal infections andopportunistic infections.

I claim:
 1. A method for treating a patient in need thereof viralinfections comprising: employing a compound represented by the generalformula (I): ##STR5## wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰,R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ are the same or different and independentlyrepresent a hydrogen atom, hydroxyl group, substituted or unsubstitutedC₁ -C₇ alkyl group, substituted or unsubstituted C₂ -C₇ alkenyl group,substituted or unsubstituted C₂ -C₇ alkynyl group, substituted orunsubstituted C₃ -C₇ cycloalkyl group, substituted or unsubstituted C₄-C₇ cycloalkenyl group, substituted or unsubstituted acyl group,substituted or unsubstituted acyloxy group, substituted or unsubstitutedacyloxy-C₁ -C₇ alkyl group, substituted or unsubstituted C₂ -C₇alkoxycarbonyl group, substituted or unsubstituted C₂ -C7alkenyl-oxycarbonyl group, substituted or unsubstituted aryloxycarbonylgroup, substituted or unsubstituted C₁ -C₇ alkoxy group, substituted orunsubstituted C₂ -C₇ alkenyloxy group, substituted or unsubstituted C₂-C₇ alkynyloxy group, substituted or unsubstituted C₃ -C₇ cycloalkyloxygroup, substituted or unsubstituted C₄ -C₇ cycloalkenyloxy group,substituted or unsubstituted aryloxy group, substituted or unsubstitutedaryl-C₁ -C₇ alkyloxy group, substituted or unsubstituted aryl-C₂ -C₇alkenyloxy group, substituted or unsubstituted C₁ -C₇ alkoxy-C₁ -C₇alkyl group or substituted or unsubstituted C₁ -C₇ alkoxy-C₁ -C₇ alkoxygroup, or R² and R³ may together represent an epoxy group, R¹ and R¹⁴may together represent an epoxy group, and R⁷ and R⁸ may togetherrepresent an oxo group, or a pharmaceutically acceptable salt thereof.2. A method according to claim 1, wherein the compound represented bythe formula (I) is an aconite-alkaloid.
 3. A method according to claim1, wherein the viral infections are herpes infections.
 4. A methodaccording to claim 1, wherein the viral infections are acquiredimmunodeficiency syndromes.
 5. A method according to claim 1, whereinthe viral infections are cytomegalovirus infections.
 6. A methodaccording to claim 1, wherein the infections are opportunisticinfections.